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Tracking Antigen-Experienced Effector T Cells In Vitro and In Vivo

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T-Cell Trafficking

Abstract

The TCR complex is a multisubunit complex, comprising at least eight transmembrane units. The clonotypic TCR α and β chains are responsible for antigen recognition, whilst the invariant chains of the CD3 complex (δ, ɛ and γ) and two zeta (ζ) polypeptides couple antigen recognition to downstream signal transduction pathways. TCRζ (CD247) functions as an amplification module in the TCR signalling cascade and is also essential for the assembly and surface expression of the TCR/CD3 complex. Loss of TCRζ expression is common in chronic infectious and inflammatory diseases, as well as in cancer. Previous work has indicated that TCRζlow-expressing cells phenotypically resemble antigen-experienced effector T cells. Here, we describe the derivation of a flow cytometry-based TCRζ expression index for the purpose of more precisely defining TCRζ expression, in addition to utilising a simple transmigration assay in the demonstration that TCRζdim T cells have intrinsic migratory properties that may explain their accumulation at sites of inflammation.

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Notes

  1. 1.

    Clone 6B10.2 is an alternative anti-TCRζ-PE antibody, which recognises a different cytoplasmic domain epitope.

References

  1. Meuer SC, Cooper DA, et al. (1983) “Identification of the receptor for antigen and major histocompatibility complex on human inducer T lymphocytes”. Science 222(4629), 1239–42.

    Article  PubMed  CAS  Google Scholar 

  2. Baniyash M. (2004) “TCR zeta-chain downregulation: curtailing an excessive inflammatory immune response”. Nat Rev Immunol 4(9), 675–87.

    Article  PubMed  CAS  Google Scholar 

  3. Irving BA, Weiss A. (1991) “The cytoplasmic domain of the T cell receptor zeta chain is sufficient to couple to receptor-associated signal transduction pathways”. Cell 64(5), 891–901.

    Article  PubMed  CAS  Google Scholar 

  4. Ohno H, Aoe T, et al. (1993) “Developmental and functional impairment of T cells in mice lacking CD3 zeta chains”. EMBO J 12(11), 4357–66.

    PubMed  CAS  Google Scholar 

  5. Yamazaki T, Arase H, et al. (1997) “A shift from negative to positive selection of autoreactive T cells by the reduced level of TCR signal in TCR-transgenic CD3 zeta-deficient mice”. J Immunol 158(4), 1634–40.

    PubMed  CAS  Google Scholar 

  6. Seitzer U, Kayser K, et al. (2001) “Reduced T-cell receptor CD3zeta-chain protein and sustained CD3epsilon expression at the site of mycobacterial infection”. Immunology 104(3), 269–77.

    Article  PubMed  CAS  Google Scholar 

  7. Liossis SN, Ding XZ, et al. (1998) “Altered pattern of TCR/CD3-mediated protein-tyrosyl phosphorylation in T cells from patients with systemic lupus erythematosus. Deficient expression of the T cell receptor zeta chain”. J Clin Invest 101(7), 1448–57.

    Article  PubMed  CAS  Google Scholar 

  8. Pang M, Setoyama Y, et al. (2002) “Defective expression and tyrosine phosphorylation of the T cell receptor zeta chain in peripheral blood T cells from systemic lupus erythematosus patients”. Clin Exp Immunol 129(1), 160–8.

    Article  PubMed  CAS  Google Scholar 

  9. Nambiar MP, Mitchell JP, et al. (2003) “Prevalence of T cell receptor zeta chain deficiency in systemic lupus erythematosus”. Lupus 12(1), 46–51.

    Article  PubMed  CAS  Google Scholar 

  10. Kono K, Ichihara F, et al. (1998) “Expression of signal transducing T-cell receptor zeta molecules after adoptive immunotherapy in patients with gastric and colon cancer”. Int J Cancer 78(3), 301–5.

    Article  PubMed  CAS  Google Scholar 

  11. Zhang Z, Gorman CL, Vermi AC, Monaco C, Foey A, Owen S, Amjadi P, Vallance A, McClinton C, Marelli-Berg FM, Isomaki P, Russell A, Dazzi F, Vyse TJ, Brennan FM, Cope AP. (2007 May 15) TCR{zeta}dim lymphocytes define populations of circulating effector cells that migrate to inflamed tissues. Blood 109(10), 4328–35.

    Article  PubMed  CAS  Google Scholar 

  12. Gorman CL,, Russell AI,, Zhang Z,, Cunninghame Graham D,, Cope AP,, Vyse TJ. (2008 Jan 15) Polymorphisms in the CD3Z gene influence TCRζeta expression in systemic lupus erythematosus patients and healthy controls. J Immunol 180(2), 1060–70.

    PubMed  CAS  Google Scholar 

  13. Whiteside TL. (2004) “Down-regulation of zeta-chain expression in T cells: a biomarker of prognosis in cancer?” Cancer Immunol Immunother 53(10), 865–78.

    Article  PubMed  CAS  Google Scholar 

  14. Monaco C, Andreakos E, et al. (2002) “T cell-mediated signaling to vascular endothelium: induction of cytokines, chemokines, and tissue factor”. J Leukoc Biol 71(4), 659–68.

    PubMed  CAS  Google Scholar 

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Gorman, C.L., Monaco, C., Ammiratti, E., Vermi, AC., Marelli-Berg, F.M., Cope, A.P. (2010). Tracking Antigen-Experienced Effector T Cells In Vitro and In Vivo. In: Marelli-Berg, F., Nourshargh, S. (eds) T-Cell Trafficking. Methods in Molecular Biology, vol 616. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-461-6_16

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  • DOI: https://doi.org/10.1007/978-1-60761-461-6_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-460-9

  • Online ISBN: 978-1-60761-461-6

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